Mesenchymal stem cells, due to their wide potential for differentiation into different cell types and the production of growth factors and various components of the extracellular matrix, offer many possibilities for therapeutic use. In addition to their use in tissue regeneration and in the modulation of the immune system, they can also serve as delivery systems for the treatment of cancer, as studies have shown that mesenchymal stem cells often migrate towards tumor tissue. Cytokine interleukin 12 (IL-12), due to its ability of activation natural and adaptive immunity and stimulation of the production of cytokine interferon ?, which coordinates anti-tumor defense, is an ideal candidate for tumor immunotherapy. The goal of our work was to obtain a stable transfected line of canine stem cells with non-viral methods, that adequately express IL-12 and could potentially be a part of anti-tumor therapy in dogs. We used the method of electroporation and lipofection. Using the reporter plasmid pEGFP N1, we optimized the conditions of electroporation, which were later used in electroporations with the therapeutic plasmid pcDNA3.1zeo + sccaIL12. The optimization included the electroporation parameters, the number of cells/electroporation, the amount of plasmid DNA/electroporation, and the type of electroporation buffer. We were not able to obtain stable transfected canine stem cells that express caIL-12 despite repeated attempts of electroporation and selection. However, with the qPCR method we successfully proved transient expression of caIL-12 at the mRNA level. The results of the master thesis have laid out good foundation for the transfection of canine stem cells derived from fat tissue.
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